BACKGROUND: Sarcoplasmic reticulum calcium ATPase 2a (SERCA2a) is a key protein that maintains myocardial Ca 2+ homeostasis. The present study aimed to investigate the mechanism underlying the SERCA2a-SUMOylation (small ubiquitin-like modifier) process after ischemia/reperfusion injury (I/RI) in vitro and in vivo . METHODS: Calcium transient and systolic/diastolic function of cardiomyocytes isolated from Serca2a knockout (KO) and wild-type mice with I/RI were compared. SUMO-relevant protein expression and localization were detected by quantitative real-time PCR (RT-qPCR), Western blotting, and immunofluorescence in vitro and in vivo . Serca2a-SUMOylation, infarct size, and cardiac function of Senp1 or Senp2 overexpressed/suppressed adenovirus infected cardiomyocytes, were detected by immunoprecipitation, triphenyltetrazolium chloride (TTC)-Evans blue staining, and echocardiography respectively. RESULTS: The results showed that the changes of Fura-2 fluorescence intensity and contraction amplitude of cardiomyocytes decreased in the I/RI groups and were further reduced in the Serca2a KO + I/RI groups. Senp1 and Senp2 messenger ribose nucleic acid (mRNA) and protein expression levels in vivo and in cardiomyocytes were highest at 6 h and declined at 12 h after I/RI. However, the highest levels in HL-1 cells were recorded at 12 h. Senp2 expression increased in the cytoplasm, unlike that of Senp1. Inhibition of Senp2 protein reversed the I/RI-induced Serca2a-SUMOylation decline, reduced the infarction area, and improved cardiac function, while inhibition of Senp1 protein could not restore the above indicators. CONCLUSION I/RI activated Senp1 and Senp2 protein expression, which promoted Serca2a-deSUMOylation, while inhibition of Senp2 expression reversed Serca2a-SUMOylation and improved cardiac function.
Animals ; Mice ; Calcium/metabolism* ; Cysteine Endopeptidases/metabolism* ; Myocardial Reperfusion Injury/metabolism* ; Myocardium/metabolism* ; Myocytes, Cardiac/metabolism* ; Proteins/metabolism* ; Sarcoplasmic Reticulum Calcium-Transporting ATPases/genetics*

Objective:To explore risk factors of in-stent restenosis (ISR) after vertebral artery stenting (VAS) in patients with ischemic cerebrovascular disease.Methods:Case-control studies and cohort studies on risk factors for ISR after VAS in patients with ischemic cerebrovascular disease were searched from CNKI, VIP, Wanfang, Chinese Biomedical literature Database, PubMed, Embase and Cochrane Library. The retrieval time limit was the establishment of each database until May 31, 2021. Two researchers screened the literature and extracted relevant data. Newcastle-Ottawa Scale (NOS) was used to evaluate quality of the literature and RevMan 5.3 software was used for meta-analysis.Results:A total of 30 studies were included. Meta-analysis results showed that smoking [ OR=3.76, 95% confidence interval ( CI) : 2.43-5.82, P<0.01], diabetes ( OR=2.95, 95% CI: 2.15-4.06, P<0.01) , hypertension ( OR=2.55, 95% CI: 1.50-4.34, P=0.006) , hyperlipidemia ( OR=7.12, 95% CI: 3.46-14.68, P<0.01) , coronary heart disease ( OR=3.06, 95% CI: 1.10-8.47, P=0.03) , homocysteine ( OR=2.36, 95% CI: 1.30-4.27, P=0.005) , clopidogrel drug-related gene ( CYP2C19) mutation ( OR=3.04, 95% CI: 1.63-5.68, P=0.005) , lesion vessel diameter ( OR=3.43, 95% CI: 1.30-9.05, P=0.01) , residual stenosis ( OR=6.08, 95% CI: 3.28-14.07, P<0.01) ) , stent type ( OR=2.26, 95% CI: 1.18-4.36, P=0.01) , stent length ( OR=3.52, 95% CI: 2.34-5.30, P<0.01) were associated with ISR after VAS operation in patients with ischemic cerebrovascular disease. Conclusions:Postoperative smoking, history of diabetes mellitus, history of hypertension, history of hyperlipidemia, history of coronary heart disease, high level of homocysteine, CYP2C19 mutation, shorter lesion vessel diameter, postoperative residual stenosis, use of bare metal stents and longer stent length are risk factors for ISR after VAS in patients with ischemic cerebrovascular disease. Clinical medical staff should adjust the follow-up time of postoperative patients according to risk factors and formulate individualized strategies to prevent the occurrence of ISR.

[Abstract] Objective: To investigate the effect of circular RNA FBXO11 (circFBXO11) regulating the miR-376a-3p/SNRPB (small nuclear ribonucleoprotein polypeptides B gene) axis on the proliferation and apoptosis of gastric cancer SNU-1 cells. Methods: Cancer and para-cancerous tissue samples from 30 patients with gastric cancer who underwent surgical resection were surgically resected in the Department of Oncosurgery, the Affiliated Hospital of North China University of Science and Technology from January 2018 to January 2019 were collected. The positive expression rate of SNRPB protein in gastric cancer tissues was detected by Immunohistochemical staining. The expression levels of circFBXO11, miR-376a-3p and SNRPB mRNA in gastric cancer tissues, gastric cancer cell lines (SNU-1, AGS and HS-746T) and gastric mucosal cell line GES1 were detected by qPCR. Dual luciferase reporter gene assay was used to determine the relationship between circFBXO11 and miR-376a-3p as well as between miR-376a-3p and SNRPB. The si-NC, si-circFBXO11, miR-NC, miR-376a-3p, si-SNRPB, si-circFBXO11+anti-miR-NC, si-circFBXO11+anti-miR-376a-3p, si-circFBXO11+pcDNA-NC, si-circFBXO11+pcDNA-SNRPB were transfected into gastric cancer SNU-1 cells, respectively. CCK-8 assay, Flow cytometry and WB assay were used to detect cell proliferation activity, apoptosis rate and protein expressions of SNRPB, cyclin D1 and C-caspase-3, respectively. Results: Compared with para-cancerous tissues, the expression level of circFBXO11 and the positive rate of SNRPB protein in gastric cancer tissues were significantly increased (all P<0.01), while the expression of miR-376a-3p was significantly decreased (P<0.01). Compared with GES1 cells, the expressions of circFBXO11 and SNRPB were significantly increased, while the expression of miR-376a-3p was significantly decreased (all P<0.01) in gastric cancer cells. circFBXO11 negatively regulated miR-376a-3p expression, and miR-376a-3p negatively regulated SNRPB expression. After inhibiting the expression of circFBXO11 or over-expressing miR-376a-3p or suppressing the expression of SNRPB, the proliferation viability of SNU-1 cells was decreased, and the apoptosis rate was increased (P<0.01). Either inhibiting miR-376a-3p or over-expressing SNRPB could partially reverse the effect of circFBXO11 suppression on proliferation and apoptosis of SNU-1 cells (all P<0.01). Conclusion: circFBXO11 is highly expressed in gastric cancer tissues. Inhibiting circFBXO11 inhibits the proliferation and induces apoptosis of gastric cancer cells, and the mechanism is related to the regulation of miR-376A-3p/SNRPB pathway.

Objective The study of capsaicin (CAP) on the effect and mechanism of indomethacin induced acute gastric mucosal injury in different period.Methods 80 SD rats were randomly divided into 8 groups with 10 rats in each group.The experiment was completed in two phases,and the Ⅰ period was 2 weeks,the Ⅱ period was 4 weeks.The Ⅰ period including group A1 (control group),group B1 (model group),group C1 (CAP group),group D1 (CAP + indomethacin group).The grouping method of the two periods were the same.The rats' gastric mucosa were damaged by indomethacin,and then killed the rats 4 hours later.Last,astric juice was collected to determine the total acidity of gastric acid,counted thegastric mucosal injury index,observed the gastric mucosa pathological injury,detected the expression of TRPV 1、CGRP、MDA、SOD and PGI2.Results The Ⅰ period:the gastric mucosa of group A1 and C1 had no damage.Group D1 compared with group B1,there was no significant difference in gastric mucosa injury (P ＞ 0.05),total acidity decreased significantly (P ＜ 0.05),MDA was no significant difference (P ＞ 0.05),SOD、PGI2 increased significantly (P ＜ 0.05),the expression of TRPV1、CGRP increased significantly (P ＜ 0.05).The Ⅱperiod:the gastric mucosa of group A2 and C2 had no damage.Group D2 compared with group B2,the gastric mucosa injury were significantly reduced (P ＜ 0.05),total acidity decreased significantly (P ＜ 0.05),MDA decreased significantly (P ＜ 0.05),SOD、PGI2 increased significantly (P ＜ 0.05),the expression of TRPV1、CGRP increased significantly (P ＜ 0.05).Conclusion There was no damage to the general morphology and histology of gastricmucosa in rats by intragastric CAP 1 mg/(kg· d) for 2 weeks and 4 weeks.2.It could prevent that indomethacininduced acute gastric mucosal injury in rats by pretreated with CAP 1 mg(kg· d) for 4weeks.

BACKGROUND:A large number of clinical trials have found that the number of bone marrow stem cels at the femoral neck and proximal femur in patients with osteonecrosis of the femoral head is significantly reduced, accompanied by decreased activity, which causes a significant decrease in osteogenic capacity that the necrotic bone cannot be effectively repaired after absorption, leading to the colapse of the femoral head. OBJECTIVE:To probe into the early clinical efficacy of autologous peripheral blood mononuclear cel transplantation with porous core decompression for treatment of avascular necrosis of the femoral head. METHODS:Forty-five patients with avascular necrosis of the femoral head (49 hips) were enroled in this study, and underwent autologous peripheral blood mononuclear cel transplantation with porous core decompression. After treatment, pain scores, Harris hip score, scores on the satisfaction of patients were evaluated, as wel as X-ray, CT and MRI examinations. RESULTS AND CONCLUSION:Al the patients received a folow-up visit of 11-14 months, averagely (12.5±0.6) months. During the folow-up, there were no complications and serious adverse reactions. Postoperative pain scores and Harris scores were both improved significantly compared with preoperative ones (P < 0.05). At 12 months after treatment, the excelent satisfaction rate was up to 92%. Patient’s MRI low signal region accounting for a percentage of the volume of the femoral head was decreased from (40.1±7.34)% preoperatively to (20.23±5.4)% at 6 months postoperatively, and there was a significant difference (P < 0.05). These findings indicate that autologous peripheral blood mononuclear cel transplantation with porous core decompression for treatment of avascular necrosis of the femoral head has significantly clinical effects at early stage, which can obviously reduce joint pain, improve and restore hip joint function, and delay progression of disease.

OBJECTIVETo investigate the protective mechanism of stearoyl-CoA desaturase 1 (SCD1) over-expression against the pro-apoptotic affects of palmitic acid on hepatocytes using the rat BRL cell line.

METHODSConcentration effect curves were generated using the trypan blue exclusion test to assess the death rate of BRL cells upon exposure to a dilution series of palmitic acid. The multiplicity of infection (MOI) of a lentiviral expression vector, pGC-FU-GFP, was determined for the BRL cells. Unmanipulated BRL cells were divided into two groups: the non-palmitate groups were composed of ordinary cultured cells (CON) alone, infected with lentivirus empty expression vector (negative control, NC), and infected with lentivirus overexpressing SCD1 (SCD1-LV); the palmitate groups were composed of ordinary cultured cells plus palmitate (CON+) alone, infected with lentivirus empty expression vector plus palmitate (NC+), and infected with lentivirus overexpressing SCD1 plus palmitate (SCD1-LV+). SCD1 mRNA expression was detected by real-time PCR. Propidium iodide (PI) single-staining was used to detect apoptosis and assess the cell cycle. Inter-group differences were analyzed statistically.

RESULTSThe death rate of BRL cells increased significantly after 72 h of exposure to 400 mumol/L palmitate (P less than 0.01). The MOI of pGC-FU-GFP in BRL cells was 20. The expression of SCD1 was significantly higher in the SCD1-LV and SCD1-LV+ groups than in the respective controls (vs. CON: F = 289, P less than 0.01; vs. CON+: F = 1522, P less than 0.01). Palmitate exposure led to decreased expression of SCD1 (CON+ vs. CON, F = 22, P less than 0.05 and NC+ vs. NC: F = 34, P less than 0.05). The ratio of S stage cells was similar in all non-palmitate groups (CON, NC and SCD1-LV, P = 0.137). However, there was a significant apoptotic peak and lower ratio of S stage cells in the control palmitate groups (CON+ and NC+) and the activity of cell proliferation was decreased as well. The ratio of apoptotic cells was decreased significantly in the SCD1-LV+ group compared to the CON+ group (P less than 0.01).

CONCLUSIONThe expression of SCD1 and its desaturation activity increased in BRL cells upon infection with the pGC-FU-SCD1-GFP lentiviral vector, suggesting that SCD1 over-expression can decrease palmitic acid-induced toxicity and apoptosis in hepatocytes.

Animals ; Apoptosis ; Cell Line ; Genetic Vectors ; Hepatocytes ; metabolism ; Lentivirus ; genetics ; Palmitic Acid ; toxicity ; Rats ; Stearoyl-CoA Desaturase ; metabolism

Objective To evaluate the performance of ACL TOP coagulation analyzer system in the laboratory of children′s hospital. Methods According to the documents of CLSI, the analytic characteristics including precision, accuracy, linearity, interference and carryover rate were examined; specimens from healthy children were collected and assayed to determine the reference range of Prothrombin Time (PT), Activated Partial Thromboplastin Time (APTT), Thrombin Time (TT), Fibrinogen (FIB) and D-Dimer (D-D) from children on ACL TOP. Results The within-run and between-day coefficient of variability (CV) were within an acceptable range; The accuracy deviation of PT , APTT and FIB were less than 1/2 allowed total errors; The results of determination of FIB linearity test were correlated with the results of calculation: Y = 1.002 1X-0.122, R2 =0.998 2; The extent of influence of low to middle grade of jaundice , fat and hemolysis on each test were all less than 1/2 allowed total error; The carryover rates were lower than 1.81% and within an acceptable range; The reference range of PT, APTT, TT and FIB were PT (9.1-13.1 s), APTT (24.9-42.1 s), TT (12.6-21.1 s), FIB (1.924-4.011 g/L). Conclusion The ACL TOP coagulation analyzer has good repeatability, stability, linearity and capability of anti-interference and anti-carryover.

Aim To elucidate the therapeutic effect of ginsenosides, berberine and jasminoidin after given a-lone or treatment with combination on the focal cerebral ischemia rats and study the compatibility mechanism. Methods We determined 12 endogenous amino acids in serum of rats after cerebral ischemia over 12 hours with RRLC-QQQ to evaluate the integrated role of YQJD at the dosage of 25 mg·kg-1 and 5 mg·kg-1 . Generally accepted methods were used, including be-havior test, One-Way AVONA, PLS-DA, as well as PCA to evaluate the injury induced by focal cerebral is-chemia. Results The score of neurological deficits and the level of five amino acids, namely Glu, Asp, Met, Hcy, Phe in the combination of ginsenosides, berberine and jasminoidin group in the dosage of 25 mg ·kg-1 and 5 mg·kg-1 significantly decreased (P<0. 05, P<0. 01) compared to those of model group. For another, the largest contribution group in the three principal components of PC1 , PC3 , PC4 at the dosage of 25 mg/kg and the six principal components PC1 ~PC5, PC7 in 5 mg·kg-1 was the combination of gin-senosides, berberine, jasminoidin group. Conclusions The results suggest that the efficacy of the combina-tion of ginsenosides, berberine and jasminoidin is su-perior to the combination of two or any single compo-nent, which can significantly improve the metabolic disorder of the endogenous amino acid after cerebral is-chemia. And it could be speculated that ginsenosides may play a more important role than berberine and jas-minoidin in regulating the level of amino acid metabo-lism.

OBJECTIVE: To investigate the clinical manifestations and diagnostic method of hyperparathyroidism due to parathyroid tumors and to evaluate the intra-operative detection of parathyroid hormone in surgical treatment. METHOD: Thirty-seven cases with functional parathyroid tumors from January 2003 to October 2012 were retrospectively analyzed. The clinical manifestation, examination and operation method, changes of parathyroid hormone before and after operation were collected. RESULT: All cases were definitely diagnosed before operation. The sensitivity and the positively predictive values of neck ultrasonography were 86.5% and 97.6% respectively, and the same data of Tc-99m-MIBI was 97.2% and 100.0%. The PTH levels declined by 84.9% ten minutes after tumor resecting compared with the level before operation. The serum calcium and PTH returned to normal levels and symptomatic relief occurred after operation. CONCLUSION Recurrent bone disease, long-term urinary calculus and obscure gastrointestinal symptoms were common symptoms of hyperparathyroidism due to parathyroid tumors. The neck ultrasonography and Tc-99m-MIBI were suitable for location of parathyroid tumors. Surgical operation was an effective treatment for parathyroid tumor. Intra-operative PTH assay would be able to ensure the radical excision and the operative safety for functional parathyroid tumors.
Adult ; Aged ; Female ; Humans ; Hyperparathyroidism ; etiology ; surgery ; Male ; Middle Aged ; Monitoring, Intraoperative ; Parathyroid Hormone ; blood ; Parathyroid Neoplasms ; complications ; surgery ; Retrospective Studies

Objective To investigate the clinical feature,pathologic characteristics and prognosis of non-muscle invasive urothelial bladder tumor in young and old patients.Methods From January 2000 to March 2011,the clinicial data of 48 young patients (age ≤ 40 years) with non-muscle invasive urothelial bladder tumor and 50 patients randomly selected with non-muscle invasive urothelial tumor (age ≥ 60 years) were analyzed and compared retrospectively.There were 38 male and 10 female with a median age of 35.4 years (range,18 to 40).There were 34 male and 16 female with a median age of 68.5 years (range,68.5 to 87).All patients had postoperative intravesical instillation for one year.Young patients presented with gross hematuria mostly,which were similmar with old patients.Solitary tumor were 45 cases and 40 cases,and the multiple tumors were 3 cases and 10 cases in the young and old groups,respectively.Of the young group,40 patients were treated by transurethral resection of bladder tumor,and 8 patients by partial cystectomy.Of the old group,35 patients were treated by transurethral resection of bladder tumor,and 15 patients by partial cystectomy.Results According to 2004 WHO classification of papillary urothelial tumor,lower grade tumor were more frequentto occur in young group than in old group.There was significant difference in incidence of PUNLMP between young group and old group (16/48,33.3％ and 8/50,16.0％,P ＜ 0.05).There was significant difference in incidence of high grade bladder cancer between young group and old group (7/48,14.6％ and 17/50,34.0％,P ＜0.05).The incidence of PTa tumor was 70.8％ and 44.0％ in the young and old groups,respectively (P ＜ 0.05).Median follow up was 34 months (range,6 to 132) in young group and 35 months (range,6 to 130) in old group,respectively.Five-year recurrence rate was 36.7％ and 64.3％ respectively (P ＜ 0.05).Conclusions Non-muscle invasive urothelial bladder tumor in young patients had a better prognosis than those in the old group,with lower grade and stage at diagnosisand lower recurrence rate.